NUCLEIC ACID MEDIATED ELECTRON TRANSFER

Abstract: The present invention provides for the selective covalent modification of nucleic acids with redox active moieties such as transition metal complexes. Electron donor and electron acceptor moieties are covalently bound to the ribose-phosphate backbone of a nucleic acid at predetermined positions. The resulting complexes represent a series of new derivatives that are bimolecular templates capable of transferring electrons over very large distances at extremely fast rates. These complexes possess unique structural features which enable the use of an entirely new class of bioconductors and photoactive probes.

Description: NUCLEIC ACID MEDIATED ELECTRON TRANSFER FIELD OF THE INVENTION The present invention is directed to electron transfer via nucleic acids. More particularly, the invention is directed to the site-selective modification of nucleic acids with electron transfer moieties such as redox active transition metal complexes to produce a new series of biomaterials and to methods of making and using them. The novel biomaterials of the present invention may be used as bioconductors and diagnostic probes. BACKGROUND OF THE INVENTION The present invention, in part, relates to methods for the site-selective modification of nucleic acids with redox active moieties such as transition metal complexes, the modified nucleic acids themselves, and their uses. Such modified nucleic acids are particularly useful as bioconductors and photoactive nucleic acid probes. The detection of specific nucleic acid sequences is an important tool for diagnostic medicine and molecular biology research. Gene probe assays currently play roles in identifying infectious organisms such as bacteria and viruses, in probing the expression of normal genes and identifying mutant genes such as oncogenes, in typing tissue for compatibility preceding tissue transplantation, in matching tissue or blood samples for forensic medicine, and for exploring homology among genes from different species. Ideally, a gene probe assay should be sensitive, specific and easily automatable (for a review, see Nickerson, Current Opinion in Biotechnology 4:48-51 (1993)). The requirement for sensitivity (i.e. low detection limits) has been greatly alleviated by the development of the polymerase chain reaction (PCR) and other amplification technologies which allow researchers to amplify exponentially a specific nucleic acid sequence before analysis (for a review, see Abramson et al., Current Opinion in Biotechnology, 4:41-47 (1993)). Specificity, in contrast, remains a problem in many currently available gene probe assays. The extent of molecular complementarity between probe and target defines the specificity of the interaction. Variations in the concentrations of probes, of targets and of salts in the hybridization medium, in the reaction temperature, and in the length of the probe may alter or influence the specificity of the probe/target interaction. It may be possible under some limited circumstances to distinguish targets with perfect complementarity from targets with mismatches, although this is generally very difficult using traditional technology, since small variations in the reaction conditions will alter the hybridization. New experimental techniques formismatch detection with standard probes include DNA ligation assays where single point mismatches prevent ligation and probe digestion assays in which mismatches create sites for probe cleavage. Finally, the automation of gene probe assays remains an area in which current technologies are lacking. Such assays generally rely on the hybridization of a labelled probe to a target sequence followed by the separation of the unhybridized free probe. This separation is generally achieved by gel electrophoresis or solid phase capture and washing of the target DNA, and is generally quite difficult to automate easily. The time consuming nature of these separation steps has led to two distinct avenues of development. One involves the development of high-speed, high-throughput automatable electrophoretic and other separation techniques. The other involves the development of non- separation homogeneous gene probe assays. For example, Gen-Probe Inc. , (San Diego, CA) has developed a homogeneous protection assay in which hybridized probes are protected from base hydrolysis, and thus are capable of subsequent chemiluminescence. (Okwu abua et al. Res. Microbiol. 143:183 (1992)). Unfortunately, the reliance of this approach on a che iluminescent substrate known for high background photon emission suggests this assay will not have high specificity. EPO application number 86116652.8 describes an attempt to use non-radiative energy transfer from a donor probe to an acceptor probe as a homogeneous detection scheme. However, the fluorescence energy transfer is greatly influenced by both probe topology and topography, and the DNA target itself is capable of significant energy quenching, resulting in considerable variability. Therefore there is a need for DNA probes which are specific, capable of detecting target mismatches, and capable of being incorporated into an automated system for sequence identification. As outlined above, molecular biology relies quite heavily on modified or labelled oligonucleotides for traditional gene probe assays (Oligonucleotide Synthesis: A Practical Approach. Gait et al., Ed., IRL Press: Oxford, UK, 1984; Oligonucleotides and Analogues: A Practical Approach. Ed. F. Eckstein, Oxford University Press, 1991) . As a result, several techniques currently exist for the synthesis of tailored nucleic acid molecules. Since nucleic acids do not naturally contain functional groups to which molecules of interest may easily be attached covalently, methods have been developed which allow chemical modification at either of the terminal phosphates or at the heterocyclic bases (Dreyer et al. Proc. Natl. Acad. Sci. USA, 1985, 82:968) . For example, analogues of the common deoxyribo- and ribonucleosides which contain amino groups at the 2' or 3' position of the sugar can be made using established chemical techniques. (See Imazawa et al., J. Org..Chem., 1979, 44:2039; Imazawa et al. , J. Org. Chem.43(15) :3044 (1978); Verheyden et al., J. Org. Chem. 36(2):250 (1971); Hobbs et al., J. Org. Chem. 42(4) :714 (1977)). In addition, oligonucleotides may be synthesized with 2'-5' or 3'-5' phosphoamide linkages (Beaucage et al., Tetrahedron 49(10):1925 (1992); Letsinger, J. Org. Chem., 35:3800 (1970); Sawai, Chem. Lett. 805 (1984); Oligonucleotides and Analogues: A Practical Approach, F. Eckstein, Ed. Oxford University Press (1991)). The modification of nucleic acids has been done for two general reasons: to create nonradioactive DNA markers to serve as probes, and to use chemically modified DNA to obtain site-specific cleavage. To this end, DNA may be labelled to serve as a probe by altering a nucleotide which then serves as a replacement analogue in the nick translational resynthesis of double stranded DNA. The chemically altered nucleotides may then provide reactive sites for the attachment of immunological or other labels such as biotin. (Gilliam et al., Anal. Bioche . 157:199 (1986)). Another example uses ruthenium derivatives which intercalate into DNA to produce photoluminescence under defined conditions. (Friedman et al., J. Am. Chem. Soc. 112:4960 (1990)). In the second category, there are a number of examples of compounds covalently linked to DNA which subsequently cause DNA chain cleavage. For example 1,10- phenanthroline has been coupled to single-stranded oligothymidylate via a linker which results in the cleavage of poly-dA oligonucleotides in the presence of Cu 2+ and 3-mercaptopropionic acid (Francois et al., Biochemistry 27:2272 (1988)) . Similar experiments have been done for EDTA'-Fe(II) (both for double stranded DNA (Boutorin et al., FEBS Lett. 172:43-46 (1986)) and triplex DNA (Strobel et al., Science 249:73 (1990)), porphyrin-Fe(III) (Le Doan et al. , Biochemistry 25:6736- 6739 (1986)), and 1,10-phenanthronine-Cu(I) (Chen et al., Proc. Natl. Acad. Sci USA, 83:7147 (1985)), which all result in DNA chain cleavage in the presence of a reducing agent in aerated solutions. A similar example using porphyrins resulted in DNA strand cleavage, and base oxidation or cross-linking of the DNA under very specific conditions (Le Doan et al., Nucleic Acids Res. 15:8643 (1987)). Other work has focused on chemical modification of heterocyclic bases. For example, the attachment of an inorganic coordination complex, Fe-EDTA, to a modified internal base resulted in cleavage of the DNA after hybridization in the presence of dioxygen (Dreyer et al., Proc. Natl. Acad. Sci. USA 82:968 (1985)). A ruthenium compound has been coupled successfully to an internal base in a DNA octomer, with retention of both the DNA hybridization capabilities as well as the εpectroscopic properties of the ruthenium label (Telser et al., J. Am. Chem. Soc. 111:7221 (1989)). Other experiments have successfully added two separate εpectroscopic labels to a single double-stranded DNA molecule (Telser et al., J. Am. Chem. Soc. 111:7226 (1989)). The study of electron transfer reactions in proteins and DNA has also been explored in pursuit of systems which are capable of long distance electron transfer. To this end, intramolecular electron transfer in protein-protein complexes, such as those found in photoεynthetic proteins and proteins in the respiration pathway, has been shown to take place over appreciable distances in protein interiors at biologically significant rates (see Bowler et al., Progress in Inorganic Chemistry: Bioinorganic Chemistry, Vol. 38, Ed. Stephen J. Lippard (1990) . In addition, the selective modification of metalloenzymeε with transition metals has been accomplished and techniques to monitor electron transfer in these systems developed. For example, electron transfer proteins such as cytochrome c have been modified with ruthenium through attachment at several histidines and the rate of electron transfer from the heme Fe 2+ to the bound Ru 3+ measured. The results suggest that electron transfer "tunnel" pathways may exist. (Baum, Chemical & Engineering News, February 22, 1993, pages 2023; see alεo Chang et al., J. Am. Chem. Soc. 113:7056 (1991)). In related work, the normal protein inεulation, which protectε the redox centerε of an enzyme or protein from nondiscriminatory reactions with the exterior solvent, was "wired" to transform these systems from electrical insulators into electrical conductors (Heller, Ace. Chem. Res. 23:128 (1990)). There are a few reports of photoinduced electron transfer in a DNA matrix. In these systems, the electron donors and acceptors are not covalently attached to the DNA, but randomly associated with the DNA, thus rendering the explicit elucidation and control of the donor-acceptor εystem difficult. For example, the intense fluorescence of certain quaternary diazoaromatic salts is quenched upon intercalation into DNA or upon exposure to individual mononucleotides, thus exhibiting electron donor processes within the DNA itself. (Brun et al., J. Am. Chem. Soc. 113:8153 (1991)). Another example of the difficulty of determining the electron transfer mechanism is found in work done with some photoexcitable ruthenium compounds. Early work suggested that certain ruthenium compounds either randomly intercalate into the nucleotide bases, or bind to the helix surface. (Purugganan et al., Science 241:1645 (1988)). A recent reference indicates that certain ruthenium compounds do not intercalate into the DNA (Satyanarayana et al.. Biochemistry 31(39):9319 (1992)) ; rather, they bind non-covalently to the surface of the DNA helix. In these early experiments, various electron acceptor compounds, such as cobalt, chromium or rhodium compounds were added to certain DNA-associated ruthenium electron donor compounds. (Puragganan et al., Science 241:1645 (1988); Orellana et al., Photochem. Photobiol. 499:54 (1991); Brun et al., J. Am. Chem. Soc. 113:8153 (1991); Davis, Chem.-Biol. Interactions 62:45 (1987); Tomalia et al., Ace. Chem. Res., 24:332 (1991)). Upon addition of these various electron acceptor compounds, which randomly bind non-covalently to the helix, quenching of the photoexcited state through electron transfer was detected. The rate of quenching was dependent on both the individual electron donor and acceptor as well as their concentrations, thus revealing the process as bimolecular. In one set of experimentε, the authorε poεtulate that the more mobile εurface bound donor promoteε electron tranεfer with greater efficiency than the intercalated εpecieε, and εuggeεt that the sugar-phosphate backbone of DNA, and possibly the solvent medium surrounding the DNA, play a significant role in the electron tranεport. (Purugganan et al.. Science 241:1645 (1988)). In other work, the authorε εtress the dependence of the rate on the mobility of the donor and acceptor and their local concentrations, and assign the role of the DNA to be primarily to facilitate an increase in local concentration of the donor and acceptor εpecieε on the helix. (Orellana et al., εupra) . In another experiment, an electron donor was reportedly randomly intercalated into the stack of bases of DNA, while the acceptor was randomly associated with the surface of the DNA. The rate of electron transfer quenching indicated a close contact of the donor and the acceptor, and the syεtem alεo exhibitε enhancement of the rate of electron tranεfer with the addition of salt to the medium. (Fromherz et al., J. Am. Chem. Soc. 108:5361 (1986)). In all of these experimentε, the rate of electron tranεfer for non-covalently bound donorε and acceptorε iε εeveral orderε of magnitude leεε than iε εeen in free εolution. An important stimulus for the development of long distance electron transfer syεtemε iε the creation of synthetic light harvesting syεtemε. Work to date suggests that an artificial light harvesting syεte containε an energy tranεfer complex, an energy migration complex, an electron tranεfer complex and an electron migration complex (for a topical review of thiε area, see Chemical & Engineering News, March 15, 1993, pages 38-48) . Two types of molecules have been tried: a) long organic moleculeε, εuch aε hydrocarbonε with covalently attached electron tranεfer species, or DNA, with intercalated, partially intercalated or helix associated electron transfer species, and b) synthetic polymers. The long organic molecules, while quite rigid, are influenced by a number of factors, which makes development difficult. These factors include the polarity and composition of the solvent, the orientation of the donor and acceptor groups, and the chemical character of either the covalent linkage or the association of the electron transfer species to the molecule. The creation of acceptable polymer electron transfer systems has been difficult because the available polymers are too flexible, such that several modes of transfer occur. Polymers that are sufficiently rigid often significantly interfere with the electron transfer mechanism or are quite difficult to synthesize. Thus the development of an electron transfer εyεtem which iε sufficiently rigid, haε covalently attached electron transfer species at defined intervals, is easy to syntheεize and doeε not appreciably interfere with the electron transfer mechanism would be useful in the development of artificial light harveεting εyεtems. In conclusion, the random distribution and mobility of the electron donor and acceptor pairs, coupled with potential short distanceε between the donor and acceptor, the looεe and preεumably reverεible association of the donors and acceptors, the reported dependence on solvent and broad putative electron pathways, and the diεruption of the DNA εtructure of intercalated compoundε rendering normal base pairing imposεible all serve as pronounced limitations of long range electron tranεfer in a DNA matrix. Therefore, a method for the production of rigid, covalent attachment of electron donorε and acceptorε to provide minimal perturbations of the nucleic acid structure and retention of its ability to base pair normally, is desirable. The present invention serveε to provide εuch a system, which allows the development of novel bioconductors and diagnostic probes. SUMMARY OF THE INVENTION The present invention provides for the selective modification of nucleic acids at specific sites with redox active moieties such as transition metal complexes. An electron donor and/or electron acceptor moiety are covalently bound preferably along the ribose- phoεphate backbone of the nucleic acid at predetermined poεitionε. The reεulting complexes represent a series of new derivatives that are biomolecular templates capable of transferring electrons over very large distances at extremely fast rateε. Theεe complexes posεeεε unique structural features which enable the use of an entirely new class of bioconductors and diagnostic probes. Accordingly, it is an object of the invention to provide a εingle εtranded nucleic acid which has both an electron donor moiety and an electron acceptor moiety covalently attached thereto. These moieties are attached through the ribose phoεphate or analogous backbone of the nucleic acid. The εingle εtranded nucleic acid iε capable of hybridizing to a complementary target sequence in a εingle stranded nucleic acid, and transferring electrons between the donor and acceptor. It iε a further object of the preεent invention to provide for a nucleic acid probe which can detect baεe- pair miεmatcheε. In this embodiment, the εingle stranded nucleic acid with a covalently attached electron donor and electron acceptor moiety is hybridized to a complementary target εequence in a εingle εtranded nucleic acid. When the region of hybridization contains at least one base pair mismatch, the rate of electron transfer between the donor moiety and the acceptor moiety is decreased or eliminated, aε compared to when there iε perfect complementarity between the probe and target sequence. It is an additional object of the present invention to provide a complex which contains a first εingle εtranded nucleic acid with at least one electron donor moiety and a second εingle εtranded nucleic acid with at least one electron acceptor moiety. As with the other embodiments of the present invention, the moietieε are covalently linked to the riboεe-phoεphate backbone of the nucleic acidε. In one aspect of the present invention, the first and second εingle εtranded nucleic acidε are capable of hybridizing to each other to form a double stranded nucleic acid, and of transferring electrons between the electron donor moiety and the electron acceptor moiety. In another aspect of the present invention, a target sequence in a εingle stranded nucleic acid compriseε at leaεt firεt and εecond target domainε, which are directly adjacent to one another. The first single εtranded nucleic acid hybridizeε to the firεt target domain and the εecond εingle εtranded nucleic acid hybridizeε to the εecond target domain, εuch that the firεt and εecond εingle εtranded nucleic acidε are adjacent to each other. Thiε reεulting hybridization complex iε capable of tranεferring electronε between the electron donor moiety and the electron acceptor moiety on the firεt and εecond nucleic acidε. In another aεpect of the preεent invention, a target sequence in a single εtranded nucleic acid compriεeε a firεt target domain, an intervening target domain, and a εecond target domain. The intervening target domain compriεeε one or more nucleotideε. The firεt and εecond εingle εtranded nucleic acids hybridize to the firεt and εecond target domainε. An intervening nucleic acid comprising one or more nucleotides hybridizes to the target intervening domain εuch that electrons are capable of being transferred between the electron donor moiety and the electron acceptor moiety on the firεt and second nucleic acids. The invention also provides for a method of making a εingle εtranded nucleic acid containing an electron tranεfer moiety covalently attached to the 5' terminuε of the nucleic acid. The method compriεeε incorporating a modified nucleotide into a growing nucleic acid at the 5' poεition to form a modified εingle εtranded nucleic acid. The modified single stranded nucleic acid is then hybridized with a complementary single εtranded nucleic acid to form a double εtranded nucleic acid. The double εtranded nucleic acid iε reacted with an electron transfer moiety such that the moiety is covalently attached to the modified εingle εtranded nucleic acid. The modified εingle εtranded nucleic acid containing the electron tranεfer moiety iε εeparated from the complementary unmodified single stranded nucleic acid. The present invention also provides a method for making a εingle stranded nucleic acid containing an electron tranεfer moiety covalently attached to an internal nucleotide. The method compriεeε creating a nucleotide dimer joined by a phoεphoramide bond and incorporating said nucleotide dimer into a growing nucleic acid to form a modified single stranded nucleic acid. The modified single stranded nucleic acid is then hybridized with a complementary single stranded nucleic acid to form a double εtranded nucleic acid. The double εtranded nucleic acid is reacted with an electron transfer moiety such that the moiety is covalently attached to the modified single stranded nucleic acid. The modified single stranded nucleic acid containing the electron transfer moiety is separated from the complementary unmodified single εtranded nucleic acid. Another aεpect of the preεent invention provideε a method of detecting a target εequence. The method comprises creating a single stranded nucleic acid with an electron donor moiety and an electron acceptor moiety covalently attached. The εingle εtranded nucleic acid containing the electron tranεfer moietieε iε then hybridized to the target εequence, and an electron tranεfer rate determined between the electron donor and the electron acceptor. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 illuεtrateε poεεible orientations of electron donor (EDM) and electron acceptor (EAM) moieties on a single εtranded nucleic acid. Figure 2 illustrates orientations of electron transfer moieties EDM and EAM on two adjacent εingle εtranded nucleic acids. These orientations also apply when the two probes are separated by an intervening sequence. Figure 3 illustrateε a εerieε of amino-modified nucleoεide precurεorε prior to incorporation into an oligonucleotide. Figureε 4A and 4B depict the εtructure of electron tranεfer moietieε. Figure 4A depictε the general formula of a repreεentative claεε of electron donorε and acceptorε. Figure 4B depictε a εpecific example of a ruthenium electron tranεfer moiety using biεbipyridine and i idazole aε the ligands. DETAILED DESCRIPTION Unleεε otherwiεe εtated, the term "nucleic acid" or "oligonucleotide" or grammatical equivalentε herein meanε at leaεt two nucleotideε covalently linked together. A nucleic acid of the preεent invention will generally contain phosphodiester bonds, although in some cases, as outlined below, a nucleic acid may have an analogouε backbone, comprising, for example, phosphoramide (Beaucage et al. , Tetrahedron 49(10) :1925 (1993) and referenceε therein; Letεinger, J. org. Chem. 35:3800 (1970)), phoεphorothioate, phoεphorodithioate, O-methylphophoroamidite linkageε (εee Eckεtein, Oligonucleotideε and Analogueε: A Practical Approach, Oxford Univerεity Preεε) , or peptide nucleic acid linkageε (εee Egholm, J. Am. Chem. Soc. 114:1895 (1992); Meier et al., Chem. Int. Ed. Engl. 31:1008 (1992); Nielεen, Nature, 365:566 (1993)). The nucleic acidε may be εingle εtranded or double εtranded, as specified. The nucleic acid may be DNA, RNA or a hybrid, where the nucleic acid contains any combination of deoxyribo- and ribo-nucleotideε, and any combination of uracil, adenine, thymine, cytoεine and guanine. In εome inεtanceε, e.g. in the case of an "intervening nucleic acid", the term nucleic acid refers to one or more nucleotides. The terms "electron donor moiety", "electron acceptor moiety", and "electron transfermoieties" orgrammatical equivalents herein refers to molecules capable of electron transfer under certain conditions. It is to be understood that electron donor and acceptor capabilities are relative; that is, a molecule which can lose an electron under certain experimental conditions will be able to accept an electron under different experimental conditions. Generally, electron tranεfer moieties contain transition metals as components, but not always. The term "target sequence" or grammatical equivalents herein means a nucleic acid sequence on a single εtrand of nucleic acid. The target εequence may be a portion of a gene, a regulatory εequence, genomic DNA, mRNA, or otherε. It may be any length, with the underεtanding that longer εequenceε are more εpecific. Generally εpeaking, this term will be understood by those skilled in the art. The probes of the present invention are designed to be complementary to the target sequence, εuch that hybridization of the target sequence and the probes of the present invention occurs. As outlined below, this complementarity need not be perfect; there may be any number of base pair mismatches which will interfere with hybridization between the target sequence and the εingle εtranded nucleic acidε of the preεent invention. However, if the number of mutations is so great that no hybridization can occur under even the leaεt stringent of hybridization conditions, the εequence is not a complementary target εequence. The termε "firεt target domain" and "εecond target domain" or grammatical equivalentε herein meanε two portionε of a target εequence within a nucleic acid which iε under examination. The firεt target domain may be directly adjacent to the εecond target domain, or the firεt and εecond target domainε may be εeparated by an intervening target domain. The terroε "first" and "εecond" are not meant to confer an orientation of the εequenceε with reεpect to the 5'-3' orientation of the target εequence. For example, aεsu ing a 5'-3' orientation of the complementary target sequence, the firεt target domain may be located either 5' to the εecond domain, or 3' to the εecond domain. The present invention is directed, in part, to the site- εelective modification of nucleic acidε with redox active moietieε εuch aε transition metal complexes for the preparation of a new εerieε of biomaterialε capable of long distance electron transfer through a nucleic acid matrix. The present invention provides for the precise placement of electron transfer donor and acceptor moieties at predetermined sites on a single εtranded or double stranded nucleic acid. In general, electron transfer between electron donor and acceptor moieties in a double helical nucleic acid does not occur at an appreciable rate unleεε nucleotide baεe pairing exiεtε in the εequence between the electron donor and acceptor in the double helical εtructure. Thiε differential in the rate of electron tranεfer forms the basiε of a utility of the preεent invention for uεe aε probes. In the εyεte of the preεent invention, where electron transfer moietieε are covalently bound to the backbone of a nucleic acid, the electronε putatively travel via the τr-orbitals of the stacked base pairs of the double stranded nucleic acid. The electron transfer rate is dependent on several factors, including the distance between the electron donor-acceptor pair, the free energy (ΔG) of the reaction, the reorganization energy (λ) , the contribution of the intervening medium, the orientation and electronic coupling of the donor and acceptor pair, and the hydrogen bonding between the baseε. The latter conferε a dependence on the actual nucleic acid εequence, since A-T pairs contain one leεε hydrogen bond than C-G pairε. However, this sequence dependence is overshadowed by the determination that there is a measurable difference between the rate of electron transfer within a DNA base-pair matrix, and the rate through the ribose-phosphate backbone, the solvent or other electron tunnels. This rate differential is thought to be at least several orders of magnitude, and may be as high aε four orders of magnitude greater through the stacked nucleotide baseε aε compared to other electron tranεfer pathwayε. Thus the presence of double stranded nucleic acids, for example in gene probe asεayε, can be determined by comparing the rate of electron tranεfer for the unhybridized probe with the rate for hybridized probeε. In one embodiment, the preεent invention provides for novel gene probes, which are uεeful in molecular biology and diagnostic medicine. In this embodiment, εingle εtranded nucleic acidε having a predetermined εequence and covalently attached electron donor and electron acceptor moietieε are εynthesized. The sequence is selected based upon a known target εequence, such that if hybridization to a complementary target sequence occurs in the region between the electron donor and the electron acceptor, electron transfer proceeds at an appreciable and detectable rate. Thus, the present invention has broad general use, as a new form of labelled gene probe. In addition, since detectable electron transfer in unhybridized probes is not appreciable, the probeε of the preεent invention allow detection of target εequences without the removal of unhybridized probe. Thus, the present invention is uniquely εuited to automated gene probe aεεayε. The preεent invention alεo finds use as a unique methodology for the detection of mutations in target nucleic acid εequenceε. Aε a reεult, if a εingle εtranded nucleic acid containing electron tranεfer moietieε iε hybridized to a target sequence with a mutation, the resulting perturbation of the base pairing of the nucleotides will measurably affect the electron tranεfer rate. This is the case if the mutation iε a εubstitution, insertion or deletion. Accordingly, the preεent invention provides for the detection of mutations in target sequenceε. Thuε, the preεent invention provides for extremely specific and εensitive probes, which may, in some embodiments, detect target εequences without removal of unhybridized probe. This will be useful in the generation of automated gene probe assays. In an alternate embodiment double stranded nucleic acids have covalently attached electron donor and electron acceptor moieties on opposite εtrandε. Such nucleic acidε are useful to detect εucceεsful gene amplification in polymerase chain reactionε (PCR) . For example, if one of the two PCR primerε containε a 5' terminally attached electron donor, and the other containε a 5' terminally attached electron acceptor, several rounds of PCR will generate doubly labeled double stranded fragments (occasionally referred to as "amplicons") . After appropriate photoinduction, the detection of electron transfer provides an indication of the εuccesεful amplification of the target εequence aε compared to when no amplification occurε. A particular advantage of the preεent invention iε that the εeparation of the εingle εtranded primerε from the amplified double εtranded DNA is not necesεary, as outlined above for probe sequenceε which contain electron tranεfer moietieε. In another embodiment the preεent invention provideε for double εtranded nucleic acids with covalently attached electron donor and electron acceptor moieties to serve as bioconductors or "molecular wire". The electron transport may occur over distanceε up to and in exceεε of 2δA per electron donor and acceptor pair. In addition, the rate of electron transfer is very fast, even though dependent on the distance between the electron donor and acceptor moieties. By modifying the nucleic acid in regular intervals with electron donor and/or electron acceptor moieties, it may be posεible to tranεport electronε over long diεtanceε, thus creating bioconductors. Theεe bioconductorε are uεeful in a large number of applicationε, including traditional applicationε for conductors such as mediators for electrochemical reactions and processes. In addition, these bioconductors may be useful as probes for photosynthesis reactions as well aε in the conεtruction of εynthetic light harveεting εyεtemε. The current modelε for the electron tranεfer component of an artificial light harveεting syεtem have several problems, as outlined above, including a dependence on solvent polarity and composition, and a lack of sufficient rigidity without arduous synthesiε. Thuε the preεent invention iε uεeful as both a novel form of bioconductor as well as a novel gene probe. In addition, the present invention provides a novel method for the εite εpecific addition to the riboεe- phoεphate backbone of a nucleic acid of electron donor and electron acceptor moietieε to a previously modified nucleotide. In one embodiment, the electron donor and acceptor moieties are added to the 3' and/or 5' termini of the nucleic acid. In alternative embodimentε, the electron donor and acceptor moieties are added to the backbone of one or more internal nucleotides, that is, any nucleotide which is not the 3' or 5' terminal nucleotide. In a further embodiment, the electron donor and acceptor moietieε are added to the backbone of both internal and terminal nucleotideε. In a preferred embodiment, the tranεition metal electron tranεfer moietieε are added through a procedure which utilizeε modified nucleotides, preferably amino-modified nucleotides. In this embodiment, the electron transfer moieties are added to the sugar phosphate backbone through the nitrogen group in phoεphoramide linkages. The modified nucleotides are then uεed to εite- εpecifically add a tranεition metal electron transfer moiety, either to the 3' or 5' termini of the nucleic acid, or to any internal nucleotide. Molecular mechanicε calculationε indicate that perturbationε due to the modification of the terminal nucleotides of nucleic acids are minimal and Watson- Crick base pairing is not disrupted (unpublished data using Biograf from Molecular Simulationε Inc. , San Diego, CA) . Accordingly, in one embodiment, modified nucleotideε are uεed to add an electron tranεfer moiety to the 5' terminus of a nucleic acid. In this embodiment, the 2' position of the riboεe of the deoxyribo- or ribonucleoside is modified prior to the addition of the electron transfer specieε, leaving the 3' poεition of the riboεe unmodified for subsequent chain attachment. In a preferred embodiment, an amino group is added to the 2 ' carbon of the sugar using establiεhed chemical techniqueε. (Imazawa et al., J. Org. Chem., 44:2039 (1979); Hobbε et al., J. Org. Chem. 42(4) :714 (1977); Verheyden et al. J. Org. Chem. 36(2):250 (1971)). Once the modified nucleotideε are prepared, protected and activated, they may be incorporated into a growing oligonucleotide by standard synthetic techniques (Gait, Oligonucleotide Synthesiε: A Practical Approach, IRL Preεε, Oxford, UK 1984; Eckεtein) aε the 5' terminal nucleotide. Thiε method therefore allowε the addition of a tranεition metal electron tranεfer moiety to the 5' terminuε of a nucleic acid. In an alternative embodiment, the 3 ' terminal nucleoεide iε modified in order to add a transition metal electron transfer moiety. In this embodiment, the 3' nucleoside is modified at either the 2' or 3' carbon of the ribose sugar. In a preferred embodiment, an amino group is added to the 2 ' or 3' carbon of the sugar using establiεhed chemical techniqueε (Imazawa et al. , J. Org. Chem., 44:2039 (1979); Hobbε et al., J. Org. Chem. 42(4) :714 (1977); Verheyden et al. J. Org. Chem. 36(2):250 (1971)). The above procedures are applicable to both DNA and RNA derivatives as shown in figure 3. The amino-modified nucleotides made as described above are converted to the 2 ' or 3' modified nucleotide triphosphate form using standard biochemical methods (Fraser et al., Proc. Natl. Acad. Sci. USA, 4:2671 (1973)). One or more modified nucleosides are then attached at the 3' end using standard molecular biology techniques such as with the uεe of the enzyme DNA polymerase I or terminal deoxynucleotidyltransferase (Ratliff, Terminal deoxynucleotidyltransferaεe. In The Enzymeε, Vol 14A. P.D. Boyer ed. pp 105-118. Academic Press, San Diego, CA. 1981) . In other embodiments, the transition metal electron tranεfer moiety or moietieε are added to the middle of the nucleic acid, i.e. to an internal nucleotide. Thiε may be accompliεhed in three wayε. In a preferred embodiment, an oligonucleotide iε amino- modified at the 5' terminus as described above. In thiε embodiment, oligonucleotide εynthesiε εi ply extends the 5' end from the amino- odified nucleotide using εtandard techniqueε. Thiε results in an internally amino modified oligonucleotide. In an alternate embodiment, electron transfer moieties are added to the backbone at a site other than riboεe. For example, phosphoramide rather than phosphodieεter linkages can be used as the site for transition metal modification. These transition metals serve aε the donorε and acceptorε for electron transfer reactions. While structural deviations from native phosphodiester linkages do occur and have been studied using CD and NMR (Heller, Ace. Chem. Res.23:128 (1990); Schuhmann et al. J. Am. Chem. Soc. 113:1394 (1991)), the phosphoramidite internucleotide link haε been reported to bind to complementary polynucleotideε and iε εtable (Beaucage et al., εupra, and referenceε therein; Letεinger, εupra; Sawai, supra; Jager, Biochemistry 27:7237 (1988)). In this embodiment, dimers of nucleotideε are created with phoεphoramide linkages at either the 2'-5' or 3'-5' positions. A preferred embodiment utilizes the 3 '-5' position for the phoεphoramide linkage, εuch that εtructural diεruption of the εubεequent Watson-Crick basepairing iε minimized. Theεe dimer unitε are incorporated into a growing oligonucleotide chain, aε above, at defined intervalε, aε outlined below. It εhould be noted that when uεing the above techniques for the modification of internal residueε it is posεible to create a nucleic acid that haε an electron transfer species on the next-to-last 3 ' terminal nucleotide, thuε eliminating the need for the extra steps required to produce the 3' terminally labelled nucleotide. In a further embodiment for the modification of internal residueε, 2 ' or 3' modified nucleoside triphosphateε are generated uεing the techniqueε deεcribed above for the 3' nucleotide modification. The modified nucleoεideε are inεerted internally into nucleic acid using standard molecular biological techniques for labelling DNA and RNA. Enzymes used for said labelling include DNA polymeraεeε εuch aε polymeraεe I, T4 DNA polymeraεe, T7 DNA polymerase, Taq DNA polymerase, reverse tranεcriptaεe and RNA polymeraεes εuch aε E. coli RNA polymeraεe or the RNA polymeraεeε from phageε SP6, T7 or T3 (Short Protocolε in Molecular Biology, 1992. Auεubel et al. Ed. pp 3.11-3.30). In a preferred embodiment, the electron donor and acceptor moietieε are attached to the modified nucleotide by methodε which utilize a unique protective hybridization εtep. In thiε embodiment, the modified εingle εtrand nucleic acid iε hybridized to an unmodified complementary sequence. This blocks the εiteε on the heterocyclic baεeε that are εusceptible to attack by the transition metal electron transfer species. The exposed amine or other ligand at the 2' or 3' position of the ribose, the phosphoramide linkages, or the other linkages useful in the present invention, are readily modified with a variety of transition metal complexes with techniques readily known in the art (εee for example Millet et al, in Metalε in Biological Syεtemε, Sigel et al. Ed. Vol. 27, pp 223-264, Marcell Dekker Inc. New York, 1991 and Durham, et al. in ACS Advanceε in Chemiεtry Serieε, Johnεon et al. Edε., Vol. 226, pp 180-193, American Chemical Society, Washington D.C. ; and Meade etal., J. Am. Chem. Soc. 111:4353 (1989)). After εucceεεful addition of the deεired metal complex, the modified duplex nucleic acid iε εeparated into single εtrandε uεing techniqueε well known in the art. In a preferred embodiment, εingle εtranded nucleic acidε are made which contain one electron donor moiety and one electron acceptor moiety. The electron donor and electron acceptor moietieε may be attached at either the 5' or 3 ' end of the εingle εtranded nucleic acid. Alternatively, the electron tranεfer moieties may be attached to internal nucleotides, or one to an internal nucleotide and one to a terminal nucleotide. It should be underεtood that the orientation of the electron tranεfer εpecieε with reεpect to the 5'-3' orientation of the nucleic acid iε not determinative. Thus, aε outlined in Figure 1, any combination of internal and terminal nucleotideε may be utilized in thiε embodiment. In an alternate preferred embodiment, εingle εtranded nucleic acidε with at leaεt one electron donor moiety and at leaεt one electron acceptor moiety are uεed to detect mutationε in a complementary target εequence. A mutation, whether it be a εubεtitution, inεertion or deletion of a nucleotide or nucleotideε, reεults in incorrect base pairing in a hybridized double helix of nucleic acid. Accordingly, if the path of an electron from an electron donor moiety to an electron acceptor moiety spans the region where the mismatch lies, the electron transfer will be eliminated or reduced εuch that a change in the relative rate will be εeen. Therefore, in thiε embodiment, the electron donor moiety iε attached to the nucleic acid at a 5' poεition from the mutation, and the electron acceptor moiety iε attached at a 3' position, or vice versa. In this embodiment it iε alεo poεεible to uεe an additional label on the modified εingle stranded nucleic acid to detect hybridization where there is one or more mismatches. If the complementary target nucleic acid contains a mutation, electron transfer is reduced or eliminated. To act as a control, the modified single stranded nucleic acid may be radio- or fluorescently labeled, εuch that hybridization to the target εequence may be detected, according to traditional molecular biology techniqueε. Thiε allowε for the determination that the target sequence existε but containε a εubstitution, insertion or deletion of one or more nucleotides. Alternatively, εingle εtranded nucleic acidε with at leaεt one electron donor moiety and one electron acceptor moiety which hybridize to regions with exact matches can be used as a controls for the preεence of the target εequence. It iε to be understood that the rate of electron transfer through a double stranded nucleic acid helix depends on the nucleotide diεtance between the electron donor and acceptor moietieε. Longer diεtanceε will have slower rateε, and consideration of the rates will be a parameter in the deεign of probeε and bioconductorε. Thuε, while it iε poεεible to meaεure rateε for diεtanceε in exceεε of 100 nucleotideε, a preferred embodiment haε the electron donor moiety and the electron acceptor moiety εeparated by at least 3 and no more than 100 nucleotideε. More preferably the moietieε are εeparated by 8 to 64 nucleotideε, with 15 being the most preferred distance. In addition, it should be noted that certain distances may allow the utilization of different detection systemε. For example, the εenεitivity of εome detection εyεtemε may allow the detection of extremely faεt rateε; i.e. the electron tranεfer moietieε may be very cloεe together. Other detection εyεtemε may require εlightly slower rates, and thus allow the electron tranεfer moieties to be farther apart. In an alternate embodiment, a single εtranded nucleic acid iε modified with more than one electron donor or acceptor moiety. For example, to increaεe the εignal obtained from theεe probes, or decreaεe the required detector εenεitivity, multiple εetε of electron donor- acceptor pairε may be uεed. Aε outlined above, in εome embodimentε different electron tranεfer moietieε are added to a εingle εtranded nucleic acid. For example, when an electron donor moiety and an electron acceptor moiety are to be added, or εeveral different electron donorε and electron acceptorε, the εyntheεiε of the εingle εtranded nucleic acid proceedε in εeveral εteps. First partial nucleic acid sequenceε are made, each containing a εingle electron tranεfer εpecieε, i.e. either a εingle tranεfer moiety or εeveral of the same tranεfer moietieε, uεing the techniqueε outlined above. Then theεe partial nucleic acid εequenceε are ligated together using techniqueε common in the art, εuch aε hybridization of the individual modified partial nucleic acidε to a complementary εingle εtrand, followed by ligation with a commercially available ligaεe. In a preferred embodiment, εingle εtranded nucleic acidε are made which contain one electron donor moiety or one electron acceptor moiety. The electron donor and electron acceptor moietieε are attached at either the 5' or 3' end of the single stranded nucleic acid. Alternatively, the electron transfer moiety is attached to an internal nucleotide. It is to be underεtood that different species of electron donor and acceptor moieties may be attached to a single stranded nucleic acid. Thus, more than one type of electron donor moiety or electron acceptor moiety may be added to any single εtranded nucleic acid. In a preferred embodiment, a firεt εingle εtranded nucleic acid iε made with on or more electron donor moietieε attached. A εecond εingle εtranded nucleic acid haε one or more electron acceptor moietieε attached. In thiε embodiment, the εingle stranded nucleic acids are made for use aε probeε for a complementary target εequence. In one embodiment, the complementary target εequence iε made up of a first target domain and a second target domain, where the firεt and εecond εequenceε are directly adjacent to one another. In thiε embodiment, the firεt modified εingle εtranded nucleic acid, which containε only electron donor moietieε or electron acceptor moietieε but not both, hybridizeε to the firεt target domain, and the εecond modified εingle εtranded nucleic acid, which containε only the correεponding electron tranεfer specieε, bindε to the second target domain. The relative orientation of the electron transfer εpecieε iε not important, aε outlined in Figure 2, and the preεent invention iε intended to include all poεεible orientationε. In the design of probes comprised of two εingle εtranded nucleic acids which hybridize to adjacent firεt and εecond target εequences, εeveral factorε εhould be conεidered. Theεe factors include the diεtance between the electron donor moiety and the electron acceptor moiety in the hybridized form, and the length of the individual εingle εtranded probeε. For example, it may be deεirable to εyntheεize only 5' terminally labelled probeε. In thiε caεe, the εingle εtranded nucleic acid which hybridizeε to the firεt εequence may be relatively εhort, εuch that the deεirable diεtance between the probeε may be acco pliεhed. For example, if the optimal diεtance between the electron transfer moieties is 15 nucleotides, then the firεt probe may be 15 nucleotideε long. In one aεpect of thiε embodiment, the two εingle εtranded nucleic acidε which have hybridized to the adjacent firεt and εecond target domainε are ligated together prior to the electron tranεfer reaction. Thiε may be done uεing εtandard molecular biology techniqueε utilizing a DNA ligase, such as T4 DNA ligase. In an alternative embodiment, the complementary target εequence will have a first target domain, an intervening target domain, and a second target domain. In thiε embodiment, the firεt modified εingle εtranded nucleic acid, which containε only electron donor moietieε or electron acceptor moietieε but not both, hybridizeε to the firεt target domain, and the εecond modified εingle εtranded nucleic acid, which containε only the correεponding electron tranεfer εpecieε, bindε to the εecond target domain. When an intervening εingle εtranded nucleic acid hybridizeε to the intervening target εequence, electron transfer between the donor and acceptor iε possible. The intervening sequence may be any length, and may comprise a εingle nucleotide. Its length, however, εhould take into consideration the desirable distanceε between the electron donor and acceptor moietieε on the firεt and εecond modified nucleic acidε. Intervening εequences of lengths greater than 14 are desirable, since the intervening εequence iε more likely to remain hybridized to form a double εtranded nucleic acid if longer intervening sequences are used. The presence or absence of an intervening sequence can be used to detect inεertionε and deletionε. In one aεpect of this embodiment, the first single stranded nucleic acid hybridized to the first target domain, the intervening nucleic acid hybridized to the intervening domain, and the εecond εingle εtranded nucleic acid hybridized to the εecond target domain, may be ligated together prior to the electron tranεfer reaction. Thiε may be done using standard molecular biology techniques. For example, when the nucleic acids are DNA, a DNA ligase, such as T4 DNA ligase can be used. The complementary target εingle εtranded nucleic acid of the preεent invention may take many forms. For example, the complementary target single εtranded nucleic acid εequence may be contained within a larger nucleic acid εequence, i.e. all or part of a gene or mRNA, a reεtriction fragment of a plaεmid or genomic DNA, among otherε. One εkilled in the art of molecular biology would underεtand how to conεtruct uεeful probes for a variety of target seguenceε uεing the preεent invention. In one embodiment, two εingle εtranded nucleic acidε with covalently attached electron transfer moieties have complementary sequences, such that they can hybridize together to form a bioconductor. In this embodiment, the hybridized duplex is capable of tranεferring at leaεt one electron from the electron donor moiety to the electron acceptor moiety. In a preferred embodiment, the individual εingle εtranded nucleic acidε are aligned εuch that they have blunt endε; in alternative embodimentε, the nucleic acidε are aligned εuch that the double helix haε coheεive ends. In either embodiment, it is preferred that there be uninterrupted double helix baεe-pairing between the electron donor moiety and the electron acceptor moiety, εuch that electronε may travel through the εtacked baεe pairε. In one bioconductor embodiment, the double εtranded nucleic acid haε one εingle εtrand nucleic acid which carrieε all of the electron tranεfer moietieε. In another embodiment, the electron tranεfer moietieε may be carried on either εtrand, and in any orientation. For example, one εtrand may carry only electron donorε, and the other only electron acceptorε or both εtrandε may carry both. In one embodiment, the double εtranded nucleic acid may have different electron tranεfer moieties covalently attached in a fixed orientation, to facilitate the long range transfer of electrons. This type of εyεtem takeε advantage of the fact that electron tranεfer εpecies may act as both electron donors and acceptors depending on their oxidative state. Thus, an electron donor moiety, after the losε of an electron, may act aε an electron acceptor, and vice verεa. Thuε, electron tranεfer moietieε may be εequentially oriented on either εtrand of the double εtranded nucleic acid εuch that directional tranεfer of an electron over very long diεtanceε may be acco pliεhed. For example, a double εtranded nucleic acid could contain a εingle electron donor moiety at one end and electron acceptor moietieε, of the same or different composition, throughout the molecule. A cascade effect of electron transfer could be accompliεhed in thiε manner, which may reεult in extremely long range tranεfer of electronε. The choice of the εpecific electron donor and acceptor pairε will be influenced by the type of electron tranεfer meaεurement uεed; for a review, εee Winkler et al., Chem. Rev. 92:369-379 (1992). When a long-lived excited εtate can be prepared on one of the redox εiteε, direct meaεurement of the electron tranεfer rate after photoinduction can be meaεured, using for example the flash-quench method of Chang et al., J. Amer. Chem. Soc. 113:7057 (1991). In this preferred embodiment, the excited redox site, being both a better acceptor and donor than the ground-state εpecieε, can tranεfer electronε to or from the redox partner. An advantage of thiε method iε that two electron tranεfer rateε may be meaεured: the photoinduced electron tranεfer rateε and thermal electron-hole recombination reactionε . Thuε differential rateε may be meaεured for hybridized nucleic acidε with perfect complementarity and nucleic acidε with iεmatcheε. In alternative embodimentε, neither redox εite haε a long lived excited εtate, and electron transfer measurementε depend upon bimolecular generation of a kinetic intermediate. For a review, εee Winkler et al., εupra. Thiε intermediate then relaxes to the thermodynamic product via intramolecular electron transfer using a quencher, as εeen below: D-A + hv → D-A * D-A * + Q → D-A + + Q- D-A + → D + -A D + -A + Q- - D-A + Q The upper limit of meaεurable intramolecular electron tranεfer rateε using this method is about 10 4 per second. Alternative embodimentε use the pulse-radiolytic generation of reducing or oxidizing radicals, which inject electrons into a donor or remove electrons from a donor, as reviewed in Winkler et al., supra. Electron tranεfer will be initiated uεing electrical, electrochemical, photon (including laεer) or chemical activation of the electron tranεfer moietieε. Theεe eventε are detected by changeε in tranεient absorption or by fluorescence or phosphoreεcence or chemilumineεcence of the electron tranεfer moietieε. In the preferred embodiment, electron transfer occurs after photoinduction with a laser. In this embodiment, electron donor moieties may, after donating an electron, serve as electron acceptors under certain circumstances. Similarly, electron acceptor moieties may serve as electron donorε under certain circu εtanceε. In a preferred embodiment, DNA iε modified by the addition of electron donor and electron acceptor moietieε. In an alternative embodiment, RNA iε modified. In a further embodiment, a double εtranded nucleic acid for use aε a bioconductor will contain some deoxyriboεe nucleotideε, εome riboεe nucleotideε, and a mixture of adenoεine, thymidine, cytoεine, guanine and uracil baεeε. In accordance with a further aεpect of the invention, the preferred formulations for donors and acceptorε will poεεeεs a transition metal covalently attached to a series of ligandε and further covalently attached to an amine group aε part of the riboεe ring (2' or 3' poεition) or to a nitrogen or εulfur atom aε part of a nucleotide dimer linked by a peptide bond, phoεphoramidate bond, phoεphorothioate bond, phoεphorodithioate bond or O-methyl phoεphora idate bond. A general formula is representative of a clasε of donorε and acceptors that may be employed is εhown in figure 4A. In this figure, M may be Cd, Mg, Cu, Co, Pd, Zn, Fe, Ru with the most preferred being ruthenium. The groups R 1 , R 2 , R 3 , R 4 , and R 5 may be any coordinating ligand that iε capable of covalently binding to the choεen metal and may include ligandε such aε NH 3 , pyridine, isonicotinamide, imidazole, bipyridine, and substituted derivative of bipyridine, phenanthrolines and substituted derivatives of phenanthrolines, porphyrins and εubεtituted derivativeε of the porphyrin family. The εtructure of a ruthenium electron transfer species using bisbipyridine and imidazole as the ligands iε shown in figure 4B. Specific examples of useful electron transfer complexes include, but are not limited to, those εhown in Table 1. TABLE 1 Donorε Acceptors Ru(bpy) 2 im-NH 2 -U Ru(NH 3 ) 5 -NH 2 -U Ru(bpy) 2 im-NH 2 -U Ru(NH 3 ) 4 py-NH 2 -U Ru(bpy) 2 im-NH 2 -U Ru(NH 3 ) 4 im-NH 2 -U Where: Ru ■ ruthenium bpy = bisbipyridine im = imidazole py β pyridine It is to be understood that the number of possible electron donor moieties and electron acceptor moieties is very large, and that one skilled in the art of electron transfer compounds will be able to utilize a number of compounds in the preεent invention. In an alternate embodiment, one of the electron tranεfer moietieε may be in the form of a εolid εupport εuch aε an electrode. When the other electron transfer moiety is in solution the εyεtem iε referred to aε a heterogenouε εyεtem aε compared to a homogenouε εyεtem where both electron donor and electron transfer moities are in the εame phaεe. The techniqueε uεed in thiε embodiment are analogoε to the wiring of proteins to an electrode except that the nucleic acids of the present invention are used rather than a redox protein (εee for example Gregg et al., J. Phyε. Chem. 95:5970 (1991); Heller et al., Senεorε and Actuatorε ., 13-14:180 (1993); and Piεhko et al., Anal. Chem., 63:2268 (1991)). In this embodiment, it is preferred that a redox polymer such aε a poly- (vinylpyridine) complex of Os(bpy) 2 Cl be cross-linked with an epoxide εuch aε diepoxide to form a redox- conducting epoxide cement which is capable of εtrongly binding to electrodeε made of conductive material εuch aε gold, vitreous carbon, graphite, and other conductive materials. This strong attachment is included in the definition of "covalently attached" for the purposes of this embodiment. The epoxide crosε-linking polymer is then reacted with, for example, an exposed amine, such aε the amine of an amino-modified nucleic acid deεcribed above, covalently attaching the nucleic acid to the complex, forming a "redox hydrogel" on the εurface of the electrode. In thiε embodiment, a εingle εtranded nucleic acid probe containing at leaεt one electron transfer moiety iε attached via thiε redox hydrogel to the εurface of an electrode. Hybridization of a target εequence can then be measured as a function of conductivity between the electron tranεfer moiety covalently attached to one end of the nucleic acid and the electrode at the other end. Thiε may be done uεing equipment and techniqueε well known in the art, εuch aε thoεe deεcribed in the referenceε cited above. In εimilar embodimentε, two nucleic acidε are utilized aε probeε aε deεcribed previously. For example, one nucleic acid is attached to a solid electrode, and the other, with a covalently attached electron tranεfer moiety, iε free in solution. Upon hybridization of a target sequence, the two nucleic acids are aligned such that electron tranεfer between the electron tranεfer moiety of the hybridized nucleic acid and the electrode occurε. The electron tranεfer iε detected aε outlined above, or by uεe of amperometric, potentiometric or conductometric electrochemical εenεorε uεing techniqueε well known in the art. The following exampleε εerve to more fully deεcribe the manner of uεing the above-deεcribed invention, aε well aε to εet forth the beεt modeε contemplated for carrying out various aεpectε of the invention. It is understood that these examples in no way serve to limit the true εcope of thiε invention, but rather are presented for illustrative purposeε. EXAMPLES The amino-modified monomer unitε are prepared by variation of publiεhed procedureε and are incorporated into a growing oligonucleotide by εtandard εynthetic techniqueε. The procedure is applicable to both DNA and RNA derivatives. Exa ple 1 Syntheεiε of an Oligonucleotide Duplex with Electron Transfer Moieties at the 5' Termini In thiε example an eight nucleotide double εtranded nucleic acid waε produced, with each εingle εtrand having a εingle electron tranεfer moiety covalently attached to the 5' terminal uridine nucleotide at the 2 ' carbon of the ribose sugar. Step 1: Synthesis of 5'-di(p-methoxyphenyl)methyl ether- '-(trifluoroacetamido)-2'-deoxyuridine 2 , -(trifluoroacetamido)-2'-deoxyuridine (2.0 g, 5.9 mmoles) prepared by minor modification of publiεhed procedureε (Imazawa, εupra) was repeatedly dissolved in a minimum of very dry CH 3 CN and rotary evaporated to dryneεε and then tranεferred to inert atmosphere vacuum line and further dried for a period of 1 hour. The following procedure for the εyntheεis of the material was adapted from Gait (supra) : Under positive presεure argon, the material was dissolved in freshly dried and distilled pyridine and with stirring, 0.05 equivalents (wt.) of 4-dimethylaminopyridine (DMAP) , 1.5 equivalentε of triethyla ine (TEA) and 1.2 equivalentε of 4,4'- dimethoxytrityl chloride (DMTr-Cl) were added to the reaction mixture. The progress of the reaction waε monitored by εilica gel TLC (98:2 methylene chloride:methanol, mobile phaεe) . After 30 minuteε, an additional 0.5 equivalentε each of DMTr-Cl and TEA were added and the reaction allowed to proceed for an additional three hourε. To thiε reaction mixture waε added an equal volume of water and the εolution extracted εeveral timeε with diethyl ether. The ether layerε were rotary evaporated to dryneεε, rediεεolved in a minimum amount of methylene chloride and purified by flaεh chromatography (99:1 methylene chloride:methanol, mobile phaεe) , to obtain the 5'-di(p- methoxyphenyl)methylether-2'-(trifluoroacetamido)-2'- deoxyuridine product. Step 2: 5'-2'-aminouridine-GCTACGA and 5'-2'-aminouridine-CGTAGCA 5'-di (p-methoxyphenyl)methyl ether-2'- (trifluoroacetamido)-2'-deoxyuridine waε dried under reduced pressure (glass) and disεolved in freεhly dried and diεtilled CH 3 CN and placed in a specially made conical vial and placed on an ABI DNA synthesizer. The program for the preparation of standard (i.e. unmodified) oligonucleotides was altered during the final base (amino-modified) addition to a 15-30 minute coupling time. The oligonucleotide was cleaved from the column by standard procedures and purified by C-18 reverse phase HPLC. In this manner 5'-2'-aminouridine- GCTACGA and 5'-2'-aminouridine-CGTAGCA were prepared. In addition, unmodified complementary εtrandε to both productε were made for uεe in the electron tranεfer moiety εyntheεiε below. Step 3: 5'-2'-ruthenium biεbipyridineimidazole- aminouridine-GCTACGA 5'-2'-aminouridine GCTACGAproduced in the previouε εtep was annealed to the complementary unmodified strand using standard techniqueε. All manipulations of the annealed duplex, prior to the addition of the transition metal complex were handled at 4°C. In order to insure that the DNA remained annealed during modification, the reactions were performed in 1M εalt. The 5'-amino modified duplex DNA waε diεεolved in 0.2 M HEPES, 0.8 M NaCl, pH 6.8 and repeatedly evacuated on a Schlenk line. Previouεly prepared ruthenium biεbipyridine carbonate waε diεsolved in the above buffer and oxygen was removed by repeated evacuation and purging with argon via a Schlenk line. The ruthenium complex was transferred to the DNA solution via cannulation (argon/vacuum) and the reaction allowed to proceed under positive pressure argon with stirring for 24 hours. To this reaction, 50 equivalents of imidazole was added to the flask and the reaction allowed to proceed for an additional 24 hours. The reaction mixture was removed from the vacuum line and applied to a PD-10 gel filtration column and eluted with water to remove excess ruthenium complex. The volume of the collected fractions was reduced to drynesε via a εpeed vac and the solid taken up in 0.1 M triethylammonium acetate (TEAC) pH 6.0. The duplex DNA was heated to 60°C for 15 minutes with 50% formamide to denature the duplex. The single stranded DNA was purified using a C-18 reverse phase HPLC column equiped with a diode array detector and employing a gradient from 3% to 35% acetonitrile in 0.1 M TEAC, pH 6.0. Step 4: 5'-2'-ruthenium tetraminepyridine- aminouridine-CGTAGCA 5'-aminouridine-CGTAGCA (0.3μm) was dissolved in 0.2 M HEPES, 0.8 M NaCl buffer, pH 6.8 and degassed on the vacuum line. To a 10 ml conical shaped flask equipped with a stirring bar and septum was slurried Ru(III) tetraaminepyridine chloride (10 μm) , in the same buffer. In a separate flask, Zn/Hg amalgam was prepared and dried under reduced pressure and the ruthenium(III) solution transferred (via cannulation) to the Zn/Hg amalgam. The immediate formation of a clear yellow solution (λ max = 406 n ) indicated that the reduced form of the ruthenium had been achieved and the reaction allowed to proceed for 30 minutes. This solution was transferred to the flask containing the amino-modified DNA and the reaction allowed to proceed at room temperature for 24 hours under argon. The reaction mixture waε removed from the vacuum line and a 50 fold exceεε of cobalt EDTA (Kirschner, Inorganic Synthesiε (1957) , pp 186) added to the εolution. The εolution waε applied to Sephadex G-25 gel filtration column to remove excess ruthenium complex and further purified by reverse phaεe HPLC aε deεcribed above. The two ruthenium modified nucleotideε were annealed by εtandard techniqueε and characterized (εee Example 5) . Example 2 Syntheεiε of Long DNA Duplexeε with Electron Transfer Moieties at the 5' Termini In this example, an in vitro DNA amplification technique, PCR (reviewed in Abramson et al., Curr. Op. in Biotech.4:41-47 (1993)) iε uεed to generate modified duplex DNA by polymerization of nucleotideε off modified primer εtrandε (Saiki et al.. Science 239:487 (1988)). Two oligonucleotideε 18 baεeε in length and not complementary to each other are εynthesized with amino- modification to the 2'-ribose position of the 5' nucleotides, aε in example 1. A εerieε of oligonucleotideε of increaεing lengthε starting at 40 baseε are chemically synthesized using standard chemistry. Each of the PCR templates shares a 5' εequence identical to one modified 18mer. The 3' end of the template oligonucleotide εhareε a εequence complementary to the other lδmer. PCR rapidly generateε modified duplex DNA by the catalyεiε of 5'-3' DNA εyntheεiε off of each of the modified lδmerε uεing the unmodified εtrand aε a template. One hundred nanomoleε of each of the two modified lδmerε are mixed in 1 ml of an aqueouε εolution containing 2,000 units of Taq polymerase, deoxyribonucleoside triphosphateε at 0.2 M each, 50 mM KC1, 10 mM Triε-Cl, pH β.δ, 1.5 mM MgCl 2 , 3 mM dithiothreitol and 0.1 mg/ml bovine εerum albumin. One femtomole of the template εtrand 40 baεeε in length iε added to the mixture. The εample is heated at 94°C for one minute for denaturation, two minutes at 55°C for annealing and three minutes at 72°C for extenεion. Thiε cycle iε repeated 30 timeε uεing an automated thermal cycler. The amplified template εequenceε with tranεition metal complexeε on both 5' termini are purified by agarose gel electrophoresiε and uεed directly in electron tranεfer applicationε. Example 3 Syntheεiε of Covalently Bound Electron Tranεfer Moieties at Internueleotide Linkages of Duplex DNA In this example, alternative backbones to phophodieεter linkages of oligonucleotides are employed. Functional groups incorporated into these internueleotide linkages serve aε the εite for covalent attachment of the electron tranεfer moietieε. Theεe alternate internueleotide linkageε include, but are not limited to, peptide bondε, phoεphoramidate bondε, phoεphorothioate bondε, phoεphorodithioate bondε and O- methylphoεphoramidate bondε. The preparation of peptide nucleic acid (PNA) followε literature procedureε (See Engholm, εupra) , with the εyntheεiε of Boc-protected pentaflurophenyl eεter of the chosen base (thymidine) . The resulting PNA may be prepared employing Merrifield's εolid-phaεe approach (Merrifield, Science, 232:341 (1986)), using a εingle coupling protocol with 0.1 M of the thiminyl monomer in 30% (v/v) DMF in CH 2 C1 2 . The progress of the reaction iε followed by quantiative ninhydrin analyεiε (Sarin, Anal. Biochem., 117:147 (1981)). The reεulting PNA may be modified with an appropriate tranεition metal complex aε outlined in example 1. The εyntheεiε of phoεphoramidate (Beaucage, εupra, Letεinger, εupra, Sawai, εupra) and N- alkylphoεphoramidateε (Jager, supra) internueleotide linkages follows standard literature procedures with only slight modification (the procedures are halted after the addition of a single baεe to the εolid εupport and then cleaved to obtain a dinucleotide phoεphoramidate) . A typical example iε the preparation of the phenyl eεter of 5'0-iεobutyloxycarbonylthymidyl- (3'-5')-5 , -amino-5'-deoxythymidine (Letεinger, J. Org. Chem. , εupra) . The dimer unitε are εubεtituted for standard oligonucleotideε at choεen intervalε during the preparation of DNA uεing eεtabliεhed automated techniqueε. Transition metal modification of the modified linkages takes place aε deεcribed in Example 1. The εyntheεiε of phoεphorothioate and phoεphorodithioate (Eckεtein, εupra, and referenceε within) internueleotide linkageε iε well documented. A publiεhed protocol utilizeε an Applied Bioεyεtemε DNA εyntheεizer uεing a modified β-cyanoethylphoεphoramidite cycle that capε after sulphurization with tetraethylthiuram diεulfide (TETD) (Iyer, J. Org. Chem. 55:4693 (1990)). The phoεphorothioate and phosphorodithioate analogs are prepared as dimers and cleaved from the εolid εupport and purified by HPLC (acetonitrile/triethylammonium acetate mobile phaεe) . Example 4 Syntheεiε of Two Oligonucleotideε Each with an Electron Transfer .oietv at the 5' Terminus In this example, two oligonucleotideε are made which hybridize to a εingle target sequence, without intervening εequences. One oligonucleotide has an electron donor moiety covalently attached to the 5' terminuε, and the other haε an electron acceptor moiety covalently attached to the 5' terminuε. In thiε example, the electron transfer species are attached via a uradine nucleotide, but one skilled in the art will understand the preεent methodε can be uεed to modify any of the nucleotideε. In addition, one εkilled in the art will recognize that the procedure iε not limited to the generation of δ-merε, but iε uεeful in the generation of oligonucleotide probes of varying lengths. The procedure is exactly aε in Example 1, except that the 8-merε generated are not complementary to each other, and instead are complementary to a target sequence of 16 nucleotides. Thus the final annealing step of step 4 of Example 1 iε not done. Inεtead, the two modified oligonucleotideε are annealed to the target εequence, and the reεulting complex iε characterized aε in Example 5. Example 5 Characterization of Modified Nucleic Acids Enzymatic digestion The modified oligonucleotides of example 1 were subjected to enzymatic digestion using establiεhed protocolε and converted to their constituent nucleoεideε by sequential reaction with phoεphodieεteraεe and alkaline phoεphataεe. By compariεon of the experimentally obtained integrated HPLC profileε and UV- viε εpectra of the digeεted oligonucleotideε to standards (including 2'-aminouridine and 2'- aminoadenine) , the presence of the amino-modified baεe at the predicted retention time and characteriεtic UV- viε spectra was confirmed. An identical procedure was carried out on the tranεition metal modified duplex DNA and aεεignmentε of conεtituent nucleoεideε demonεtrated single-site modification at the predicted site. Fluorescent labeled amino-modified oligonucleotides It has been demonstrated that the fluorochrome, fluoreεcein iεothiocyanate (FITC) iε εpecific for labeling primary amineε on modified oligonucleotideε while not bonding to amineε or amideε preεent on nucleotide baεeε (Haugland, Handbood of Fluoreεcent Probeε and Research Chemicals, 5th Edition, (1992)). Thiε reaction waε carried out uεing the amino- oligonucleotide εyntheεized aε deεcribed in example 1 and on an identical baεeε εequence without the 2'-amino- riboεe group preεent. Fluoreεcence εpectroεcopic meaεurementε were acquired on both theεe oligonucleotideε and the reεultε confirm the preεence of the amine on the 5'-terminal riboεe ring. Thermodvnamic Melting Curves of Modified Duplex DNA Awell establiεhed technique for meaεuring thermodynamic parameterε of duplex DNA iε the acquiεition of DNA melting curveε. A serieε of melting curves as a function of concentration of the modified duplex DNA was measured via temperature controlled UV-vis (Hewlett- Packard) , using techniques well known in the art. These results confirm that hybridization of the amino-modified and transition metal modified DNA had taken place. In addition, the results indicate that the modified DNA form a stable duplex comparable to the stability of unmodified oligonucleotide standards. Two Dimensional Nuclear Magnetic Resonance (NMR) Spectroscopy The amino-modified oligonucleotides synthesized as a part of this work were prepared in sufficient quantities (6 micromoles) to permit the assignment of the H proton NMR spectra using a 600MHz Varian NMR spectrometer. Measurement of the rate of electron transfer An excellent review of the measurement techniques is found in Winkler et al., Chem. Rev. 92:369-379 (1992). The donor is Ru(bpy) 2 (NHuridine) im, E°~l V, and the acceptor is Ru(NH 3 ) 4 py(NHuridine) im, E°~330 mV. The purified transition metal modified oligonucleotides (U MHRu<b P y)2i_P CATCGA anα U NHRu(NH3)4( P y)i CGATGCA were annealed by heating an equal molar mixture of the oligonucleotides (30 μmolar: 60 nmoles of DNA in 2 ml buffer) in pH 6.8 (100 mM NaPi, 900 mM NaCl) to 60°C for 10 minutes and slowly cooling to room temperature over a period of 4 hours. The solution was transferred to an inert atmosphere cuvette equipped with adapters for attachment to a vacuum line and a magnetic stirring bar. The solution was degassed εeveral times and the sealed apparatus refilled repeatedly with Ar gas. The entire apparatus was inserted into a cuvette holder as part of the set-up using the XeCl excimer-pumped dye laεer and data acquired at εeveral wavelengthε including 360, 410, 460 and 480 nm. The photoinduced electron tranεfer rate iε 1.6 X 10 6 ε "1 over a distance of 28 A. Example 6 Synthesiε of a Single Stranded Nucleic Acid Labeled with Two Electron Tranεfer Moietieε Thiε example uεeε the baεic procedureε deεcribed earlier to generate two modified oligonucleotideε each with an electron tranεfer moiety attached. Ligation of the two modified strands to each other produces a doubly labeled nucleic acid with any of four configurations: 5' and 3' labeled termini, 5' labeled terminuε and internal nucleotide label, 3' labeled terminus and internal nucleotide label, and double internal nucleotide labels. Specifically, the εyntheεiε of an oligonucleotide 24 baεeε in length with an electron tranεfer donor moiety on the 5' end and an internal electron tranεfer moiety iε deεcribed. Five hundred nanomoleε of each of two 5'-labeled oligonucleotideε 12 baεes in length are εyntheεized aε detailed above with ruthenium (II) biεbipyridine imidazole on one oligonucleotide, "D" and ruthenium (III) tetraamine pyridine on a εecond oligonucleotide, "A". An unmodified oligonucleotide 24 baεeε in length and complementary to the juxtapoεition of oligonucleotide "D" followed in the 5' to 3' direction by oligonucleotide "A" iε produced by εtandard εynthetic techniqueε. Five hundred nanomoleε of thiε hybridization template iε added to a mixture of oligonucleotideε "A" and "D" in 5 ml of an aqueouε εolution containing 500 mM Triε-Cl, pH 7.5, 50 mM MgCl 2 , 50 mM dithiothreitol and 5 mg/ml gelatin. To promote maximal hybridization of labeled oligonucleotides to the complementary εtrand, the mixture iε incubated at 60°C for 10 minuteε then cooled εlowly at a rate of approximately 10°C per hour to a final temperature of 12°C. The enzymatic ligation of the two labeled εtrandε iε achieved with T4 DNA ligaεe at 12°C to prevent the ligation and oligomerization of the duplexed DNA to other duplexeε (blunt end ligation) . Alternatively, E__. coli DNA ligaεe can be uεed aε it doeε not catalyze blunt end ligation. One hundred Weiεε units of T4 DNA ligaεe iε added to the annealed DNA and adenosine triphoεphate is added to a final concentration of 0.5 mM. The reaction which catalyzeε the formation of a phoεphodieεter linkage between the 5' terminal phoεphate of oligonucleotide "A" and the 3' terminal hydroxyl group of oligonucleotide "D" iε allowed to proceed for 18 hours at 12°C. The reaction is terminated by heat inactivation of the enzyme at 75 β C for 10 minuteε. The doubly labeled oligonucleotide iε εeparated from the εingly labeled oligonucleotideε and the complementary unlabeled oligonucleotide by HPLC in the preεence of urea aε in the previouε exa pleε. The doubly labeled oligonucleotide of thiε example iε ideally εuited for uεe aε a photoactive gene probe aε detailed below. Example 7 ϋεe of a Doubly Modified Oligonucleotide with Electron Tranεfer Moietieε aε a Photoactive Probe for Detection of Complementary Nucleic Acid Sequence Thiε example utilizes the oligonucleotide 24mer of example 6 in a unique type of gene-probe asεay in which removal of unhybridized probe prior to signal detection is not required. In the aεεay procedure, a region of the gag gene of human immunodeficiency viruε type I (HIV-I) iε amplified by the polymeraεe chain reaction (Saikietal., Science 239:487-491 (198δ)). Thiε region of HIV-I iε highly conεerved among different clinical iεolateε. The amplified target DNA verεuε controlε lacking in HIV- I DNA are added to a hybridization εolution of 6XSSC (0.9 M NaCl, 0.09 M Na citrate, pH 7.2) containing 50 nano oles of doubly labeled 24mer probe of example 6. Hybridization iε allowed to proceed at 60°C for 10 minuteε with gentle agitation. Detection of electron tranεfer following laεer excitation iε carried out aε in example 5. Control εampleε which lack the hybridized probe εhow negligible electron tranεfer rateε. Probeε hybridized to the gag εequence εhow efficient and rapid electron tranεfer through the DNA double helix, providing a highly εpecific, ho ogeneouε and automatable HIV-I detection aεεay. A εimilar homogeneous gene probe asεay involveε the uεe of two probeε, one an electron donor and the other an electron acceptor, which hybridize with the gagr region of HIV-I in a tandem configuration, one probe abutting the other. In thiε aεεay, electronic coupling between the two electron tranεfer moietieε dependε entirely on hybridization with the target DNA. If appropriate, the electron tranεfer from one probe to the other iε enhanced by the ligation of the juxtapoεed endε using T4 DNA ligase aε in example 6.

Claims: HAT I S CLAIMED I S : 1. A single-εtranded nucleic acid containing at leaεt one electron donor moiety and at leaεt one electron acceptor moiety, εaid electron donor moiety and εaid electron acceptor moiety being covalently attached to εaid nucleic acid. 2. A εingle-εtranded nucleic acid according to claim 1 wherein εaid covalent attachment iε to the riboεe- phoεphate backbone of εaid nucleic acid. 3. A εingle εtranded nucleic acid according to claim 1, wherein εaid εingle εtranded nucleic acid iε capable of hybridizing to a complementary target εequence in a εecond εingle εtranded nucleic acid to form a hybridization complex. 4. A εingle εtranded nucleic acid according to claim 3. wherein εaid hybridization complex iε capable of tranεferring at leaεt one electron between εaid electron donor moiety and εaid electron acceptor moiety. 5. A compoεition compriεing a firεt εingle εtranded nucleic acid containing at least one electron donor moiety and a second εingle εtranded nucleic acid containing at leaεt one electron acceptor moiety, wherein εaid electron donor moiety and electron acceptor moiety are covalently linked to the riboεe-phoεphate backbone of εaid firεt and εecond εingle εtranded nucleic acidε. 6. A compoεition according to claim 5 wherein εaid firεt εingle stranded nucleic acid is capable of hybridizing to εaid εecond εingle εtranded nucleic acid to form a double εtranded nucleic acid. 7. A double stranded nucleic acid composition according to claim 6 wherein said firεt εingle εtranded nucleic acid iε hybridized to εaid εecond εingle εtranded nucleic acid. 8. A compoεition according to claim 7 wherein εaid compoεition iε capable of tranεferring at leaεt one electron between εaid electron donor moiety and εaid electron acceptor moiety. 9. A compoεition according to claim 5 wherein said first and second εingle stranded nucleic acids are capable of hybridizing to a target sequence stranded nucleic acid, comprising at least a firεt target domain and a second target domain, wherein said first nucleic acid is capable of hybridizing to said first target domain and said second nucleic acid is capable of hybridizing to said εecond target domain to form a hybridization complex. 10. A compoεition according to claim 9 wherein εaid firεt target domain and εaid εecond target domain are adjacent to one another. 11. A composition according to claim 10 wherein said first nucleic acid and said εecond nucleic acid in εaid hybridization complex are ligated together. 12. A compoεition according to claim 10 wherein εaid hybridization complex iε capable of transferring at least one electron between εaid electron donor moiety and said electron acceptor moiety. 13. A composition according to claim 10 wherein said target sequence further comprises an intervening target domain between said first and εaid εecond target domain. 14. A composition according to claim 13 further comprising an intervening single εtranded nucleic acid εequence capable of hybridizing to said intervening target domain to form a hybridization complex. 15. A composition according to claim 14 wherein said hybridization complex is capable of transferring at least one electron between εaid electron donor moiety and εaid electron acceptor moiety. 16. A method for making a εingle εtranded nucleic acid containing an electron tranεfer moiety at the 5' terminuε, co priεing a) incorporating a modified nucleotide into a growing nucleic acid at the 5' position to form a modified single εtranded nucleic acid; b) hybridizing εaid modified εingle εtranded nucleic acid with a complementary εingle εtranded nucleic acid to form a double εtranded nucleic acid; c) reacting εaid double εtranded nucleic acid with an electron transfer moiety εuch that εaid moiety iε covalently attached to εaid modified εingle εtranded nucleic acid; and d) εeparating εaid complementary εingle stranded nucleic acid from εaid modified εingle εtranded nucleic acid containing εaid electron transfer moiety. 17. A method for making a single stranded nucleic acid containing an electron transfer moiety covalently attached to an internal nucleotide, comprising a) incorporating a modified nucleotide dimer into a growing nucleic acid to form a modified εingle εtranded nucleic acid; b) hybridizing εaid modified εingle εtranded nucleic acid with a complementary εingle εtranded nucleic acid to form a double εtranded nucleic acid; c) reacting εaid double εtranded nucleic acid with an electron transfer moiety such that εaid moiety iε covalently attached to εaid modified εingle εtranded nucleic acid; and d) εeparating εaid complementary εingle εtranded nucleic acid from the modified εingle εtranded nucleic acid containing εaid electron tranεfer moiety. 18. A method for making a εingle εtranded nucleic acid containing an electron tranεfer moiety covalently attached to the 3' terminal nucleotide, compriεing a) incorporating a modified nucleotide via enzymatic addition or replacement into a nucleic acid; b) hybridizing εaid modified εingle εtranded nucleic acid with a complementary εingle εtranded nucleic acid to form a double εtranded nucleic acid; c) reacting εaid double εtranded nucleic acid with an electron tranεfer moiety εuch that εaid moiety iε covalently attached through said phosphoramide bond of said modified εingle εtranded nucleic acid; and d) εeparating εaid complementary εingle εtranded nucleic acid from the modified εingle εtranded nucleic acid containing εaid electron tranεfer moiety. 19. A method of detecting a target εequence in a nucleic acid εample compriεing a) hybridizing a εingle εtranded nucleic acid containing at leaεt one covalently attached electron donor moiety and at leaεt one covalently attached electron acceptor moiety to εaid target εequence to form a hybridization complex; b) determining the electron transfer rate between εaid electron donor moiety and said electron acceptor moiety in the hybridization complex; and c) comparing said electron transfer rate with the electron transfer rate in the absence of the target εequence as an indicator of the presence or absence of εaid target εequence. 20. A method of detecting a target εequence in a nucleic acid wherein εaid target sequence compriseε a firεt target domain and a εecond target domain adjacent to εaid firεt target domain, wherein εaid method compriεeε: a) hybridizing a firεt nucleic acid containing at leaεt one electron donor moiety to εaid firεt target domain; b) hybridizing a εecond nucleic acid containing at leaεt one electron acceptor moiety to said εecond target domain; c) determining the electron transfer rate between εaid electron donor moiety and εaid electron acceptor moiety while εaid firεt and εecond nucleic acidε are hybridized to εaid firεt and εecond target domainε; and d) comparing εaid electron tranεfer rate with the electron tranεfer rate in the abεence of the target εequence aε an indicator of the preεence or absence of εaid target εequence in εaid nucleic acid εample.

Inventors: FRASER SCOTT E, KAYYEM JON F, MEADE THOMAS J, FRASER, SCOTT, E., KAYYEM, JON, F., MEADE, THOMAS, J.
Applicants: CALIFORNIA INST OF TECHN, CALIFORNIA INSTITUTE OF TECHNOLOGY
Application year: 1994
Main classification: C07H21
IPC codes: C07H21, C07H23, C12N15, C12Q1, G01N33